U.S. patent application number 12/094727 was filed with the patent office on 2010-01-07 for biosurfactant-containing skin care cosmetic and skin roughness-improving agent.
This patent application is currently assigned to TOYO BOSEKI KABUSHIKI KAISHA. Invention is credited to Tokuma Fukuoka, Tomohiro Imura, Masaru Kitagawa, Dai Kitamoto, Tomotake Morita, Atsushi Sogabe, Michiko Suzuki, Shuhei Yamamoto.
Application Number | 20100004472 12/094727 |
Document ID | / |
Family ID | 38067182 |
Filed Date | 2010-01-07 |
United States Patent
Application |
20100004472 |
Kind Code |
A1 |
Kitagawa; Masaru ; et
al. |
January 7, 2010 |
BIOSURFACTANT-CONTAINING SKIN CARE COSMETIC AND SKIN
ROUGHNESS-IMPROVING AGENT
Abstract
The present invention relates to a cosmetic for skin roughness
improvement/skin care containing a biosurfactant, particularly
MEL-A, MEL-B or MEL-C.
Inventors: |
Kitagawa; Masaru; (Shiga,
JP) ; Suzuki; Michiko; (Shiga, JP) ; Yamamoto;
Shuhei; (Shiga, JP) ; Sogabe; Atsushi; (Shiga,
JP) ; Kitamoto; Dai; (Ibaraki, JP) ; Imura;
Tomohiro; (Ibaraki, JP) ; Fukuoka; Tokuma;
(Ibaraki, JP) ; Morita; Tomotake; (Ibaraki,
JP) |
Correspondence
Address: |
FISH & RICHARDSON P.C.
P.O. BOX 1022
MINNEAPOLIS
MN
55440-1022
US
|
Assignee: |
TOYO BOSEKI KABUSHIKI
KAISHA
Osaka
JP
NATIONAL INSTITUTE OF ADVANCED INDUSTRIAL SCIENCE AND
TECHNOLOGY
Tokyo
JP
|
Family ID: |
38067182 |
Appl. No.: |
12/094727 |
Filed: |
November 21, 2006 |
PCT Filed: |
November 21, 2006 |
PCT NO: |
PCT/JP2006/323239 |
371 Date: |
May 22, 2008 |
Current U.S.
Class: |
549/417 |
Current CPC
Class: |
A61K 8/602 20130101;
A61Q 19/00 20130101; A61Q 19/008 20130101 |
Class at
Publication: |
549/417 |
International
Class: |
C07D 309/10 20060101
C07D309/10 |
Foreign Application Data
Date |
Code |
Application Number |
Nov 25, 2005 |
JP |
2005-340902 |
Jun 22, 2006 |
JP |
2006-172238 |
Claims
1. A skin care cosmetic comprising at least one biosurfactant.
2. The skin care cosmetic according to claim 1, for improving skin
roughness.
3. The skin care cosmetic according to claim 1, for improving skin
roughness by an action of a surfactant.
4. The skin care cosmetic according to claim 1, wherein the
biosurfactant is a mannosylerythritol lipid (MEL) and/or a
mannosylmannitol lipid (MML).
5. The skin care cosmetic according to claim 1, wherein the
biosurfactant is at least one selected from the group consisting of
mannosylerythritol lipid A (MEL-A), mannosylerythritol lipid B
(MEL-B) and mannosylerythritol lipid C (MEL-C).
6. The skin care cosmetic according to claim 1, wherein the
biosurfactant is mannosylerythritol lipid B (MEL-B).
7. The skin care cosmetic according to claim 1, wherein the
biosurfactant is mannosylerythritol lipid C (MEL-C).
8. The skin care cosmetic according to claim 1, wherein the
biosurfactant is mannosylerythritol lipid A (MEL-A).
9. A skin roughness-improving agent composed of at least one
biosurfactant.
10. A skin care cosmetic according to claim 4, wherein the
biosurfactant is MEL produced by yeast belonging to Pseudozyma
sp.
11. A skin care cosmetic according to claim 10, wherein the yeast
is Pseudozyma tsukubaensis.
Description
TECHNICAL FIELD
[0001] The present invention relates to the use of a biosurfactant
or a premixed product thereof for skin care/skin roughness
improvement, in particular the use of a biosurfactant as a
cosmetic, and further skin care/skin roughness-improvement
cosmetics containing the biosurfactant or the premixed product
thereof. More specifically, the present invention relates to a
cosmetic characterized in that the biosurfactant is a
mannosylerythritol lipid (hereinafter referred to as a "MEL"),
e.g., mannosylerythritol lipid A (hereinafter referred to as
"MEL-A"), mannosylerythritol lipid B (hereinafter referred to as
"MEL-B") or mannosylerythritol lipid C (hereinafter referred to as
"MEL-C"); or a mannosylmannitol lipid (hereinafter referred to as a
"MML"). In addition, the present invention relates to a skin
roughness-improving agent.
BACKGROUND ART
[0002] Rough skin refers to skin in a dry state, on which the
exfoliation of corneocytes is observed. This type of rough skin
develops due to an elution of intercorneocyte lipids such as
cholesterol, ceramide and fatty acids, the formation failure of a
horny layer permeation barrier because of the denaturation of the
corneocytes, and the disturbance of the
proliferation/keratinization balance in epidermal cells
attributable to ultraviolet light and detergents. Supplying
intercorneocyte lipid components or synthetic intercorneocyte
lipids analogous thereto for the purpose of preventing or treating
aforementioned rough skin has been studied.
[0003] Lamella granules biosynthesized in cells in a spinous layer
and a granular layer are released in intercellular space just under
the horny layer, extend to take a lamella structure and spread in
the intercellular space to produce the aforementioned
intercorneocyte lipids. The lamella granules are composed of
glycosylceramide, cholesterol, ceramide, phospholipids and the
like; however, glycosylceramide is rarely included in the
intercorneocyte lipids. That is, it is believed that
glycosylceramide in the lamella granules is hydrolyzed with
.beta.-glucocerebrosidase and converted into ceramide, and that
this ceramide takes the lamella structure, resulting in improved
formation of the horny layer permeation barrier as the
intercorneocyte lipid acts as a barrier to prevent rough skin. It
has been reported that a ceramide supplement is effective against
skin roughness due to a washing agent, and is highly effective for
the improvement of skin roughness (Non-Patent Literature 1).
[0004] An extraction solution from a plant is composed mainly of
glycosylceramide, but is not yet a satisfactory alternative to
ceramide. During its synthesis, there are many reaction steps, and
the cost to produce it on a large scale is high.
[0005] MEL is a yeast-produced, natural surfactant whose various
physiological actions have been previously reported (Non-Patent
Literature 2). Mannosylmannitol lipids (MML) obtained by replacing
erythritol with mannitol have been discovered recently (Patent
Document 1). Their use in external preparations and cosmetics, as
anti-inflammatory agents and anti-allergy agents (Patent Document
2), as baldness remedies and hair growth drugs (Patent Document 3),
as well as their antibacterial actions (Patent Document 4) and
surface tension lowering actions (Patent Document 5) have been
recognized; however, the use of MEL as an alternative to ceramide
effective for improving skin roughness has been unknown.
[0006] Patent Document 1: JP 2005-104837-A
Patent Document 2: JP 2005-68015-A
Patent Document 3: JP 2003-261424-A
Patent Document 4: JP Sho-57-145896-A
Patent Document 5: JP Sho-61-205450
Non-Patent Literature 1: Hihu to Biyoh (Skin and Beauty) 36:210,
2004
Non-Patent Literature 2: Journal of Bioscience and Bioengineering
94:187, 2002
DISCLOSURE OF INVENTION
Problem to be Solved by the Invention
[0007] Ceramide is used in cosmetics as a component useful for skin
roughness, but because a synthesized product or an extracted
product made from plants is expensive, only a small amount of
ceramide is currently used. It is a major object of the present
invention to provide an external preparation for skin using a
biosurfactant produced by a microorganism as an alternative to
ceramide. That is, the present invention provides the biosurfactant
by which the formation of the horny layer permeation barrier is
improved, thereby achieving the improvement of rough skin and that
is an easily available lipid component, as an external preparation
for the skin.
Means for Solving the Problems
[0008] As a result of an extensive study to overcome the above
problems, it was found that the biosurfactant acts as an
alternative to ceramide after adding the biosurfactant to a skin
roughness model made by a three-dimensional skin model defatted
with sodium lauryl sulfate (SDS); the usefulness of the
biosurfactant was further proved by applying it on a rough skin
site produced by treating human skin with SDS. That is, it was
discovered that the biosurfactant could be used not only as an
emulsifier, but could also be used in place of ceramide; thus, the
present invention was completed.
[0009] The present invention relates to the following skin care
cosmetic and skin roughness-improving agent.
[0010] (1) A skin care cosmetic comprising at least one
biosurfactant.
[0011] (2) The skin care cosmetic according to (1) for improving
skin roughness.
[0012] (3) The skin care cosmetic according to (1) for improving
skin roughness by an action of a surfactant.
[0013] (4) The skin care cosmetic according to (1), wherein the
biosurfactant is a mannosylerythritol lipid (MEL) and/or a
mannosylmannitol lipid (MML).
[0014] (5) The skin care cosmetic according to (1), wherein the
biosurfactant is at least one selected from the group consisting of
mannosylerythritol lipid A (MEL-A), mannosylerythritol lipid B
(MEL-B) and mannosylerythritol lipid C (MEL-C).
[0015] (6) The skin care cosmetic according to (1), wherein the
biosurfactant is the mannosylerythritol lipid B (MEL-B).
[0016] (7) The skin care cosmetic according to (1), wherein the
biosurfactant is the mannosylerythritol lipid C (MEL-C).
[0017] (8) The skin care cosmetic according to (1), wherein the
biosurfactant is the mannosylerythritol lipid A (MEL-A).
[0018] (9) A skin roughness-improving agent consisting of at least
one biosurfactant.
EFFECT OF THE INVENTION
[0019] It has been found in the present invention that MELs such as
MEL-A, MEL-B and MEL-C, and MML, which are the biosurfactants
produced by microorganisms, can be used in place of ceramide for
skin care and skin roughness improvement. The biosurfactant of the
present invention can be produced on a large scale by culturing the
microorganism. By the use thereof of the ceramide alternative, skin
roughness improvement/skin care action and an emulsifying action
can be expected. Thus, it is possible to obtain an external
preparation for the skin that is effective for improving skin
roughness. In particular, MEL-B and MEL-C are highly hydrophilic,
and can make stable emulsifiers. The biosurfactant may be used as a
premixed product.
[0020] MELs are preferable because MELs may be combined in
cosmetics and external preparations for the skin by dissolution in
an oil base or in an oil-soluble component, and can be prepared as
an aqueous solution (e.g., skin lotion, moisturizing liquid) by
incorporating in the MEL a liposome, which is excellent for
incorporation into the skin. The liposome can be used in a form
other than in an aqueous solution. It is not necessary for all of
the MELs to be formed into liposomes, and MELs as liposomes may be
mixed with lamella-shaped MELs or MELs with simple bodies.
[0021] The biosurfactant of the present invention is particularly
useful as a skin care cosmetic and as a cosmetic for the
improvement of skin roughness; it is also useful as a quasi-drug
and a pharmaceutical, such as a therapeutic agent, for skin
diseases such as moderate to severe skin roughness, acne, eczema,
asteatosis, senile xeroderma and skin pruritus.
[0022] The biosurfactant is useful as the component combined with
the cosmetics for washing because the biosurfactant has a detergent
property in addition to its skin roughness improvement/skin care
action.
BRIEF DESCRIPTION OF DRAWINGS
[0023] FIG. 1 is a graph showing the effects of MEL by viability
(absorbance of formazan) in a skin roughness model made by using a
three-dimensional skin model and treatment with SDS;
[0024] FIG. 2 is a graph showing the recovery of moisture in the
horny layer when an MEL-A-containing cream is applied on skin on an
upper portion of a human arm roughened by treatment with SDS;
[0025] FIG. 3 is a graph showing the effects of MEL-A, MEL (OL) and
MEL (MY) by the viability (absorbance of formazan) in the skin
roughness model made by using the three-dimensional skin model and
treatment with SDS; MEL-A=MEL produced by culturing with soybean
oil; MEL (MY)=MEL produced by culturing with methyl myristate and
MEL-A (OL)=MEL produced by culturing with olive oil;
[0026] FIG. 4 is a graph showing the effects of MEL-B and MEL-C by
the viability (absorbance of formazan) in the skin roughness model
made by using the three-dimensional skin model and treating with
SDS; MEL-B is the increased skin roughness improvement over MEL-A;
in the figure, MEL-A (OL) is MEL-A produced using olive oil as a
raw material, MEL-B (OL) is MEL-B produced using olive oil as a raw
material, MEL-A (SB) is MEL-A produced using soybean oil as a raw
material, MEL-B (SB) is MEL-B produced using soybean oil as a raw
material, and MEL-C(SB) is MEL-C produced using soybean oil as a
raw material;
[0027] FIG. 5 is a graph showing the recovery effects of the
moisture contents in the horny layer when a MEL-B-containing cream
was applied on skin on an upper portion of a human's arm roughened
by treatment with SDS;
[0028] FIG. 6 is a graph showing the dispersion stability of MEL-B
and MEL-C. MEL-B and MEL-C are more excellent than MEL-A in water
dispersion stability; there is little observable change in
turbidity because a stable suspension state is maintained even
after 6 hours; and
[0029] FIG. 7 is a graph showing the results when a MEL-B liposome
aqueous solution and a MEL-B suspension are subjected to a skin
roughness test using the three-dimensional skin model, and the
effects are examined.
BEST MODES FOR CARRYING OUT THE INVENTION
[0030] The "biosurfactant" herein is a generic name for substances
produced by organisms and having an active surface and an
emulsification action, and exhibiting not only excellent surface
activity and high biodegradability, but also a likeliness to
express behaviors and functions different from those of the
synthetic surfactants because it has various physiological
actions.
[0031] The "premixed product" is one in which a dispersant is added
in addition to the functional material, or that is diluted with a
solvent to be easily used upon manufacturing the cosmetics. The
biosurfactant of the present invention may be provided in the form
of the premixed product, in which the dispersant and the solvent
have been mixed as a cosmetic or a cosmetic additive for skin
roughness improvement/skin care.
[0032] Ceramide is a sphingolipid that occupies about 50% of the
intercellular lipids in the horny layer. Ceramide derived from
bovine brain was often used previously, but, since the spread of
mad cow disease, ceramide derived from plants has been required for
cosmetics.
[0033] Ceramide-like actions are the actions in ceramide, the major
intercellular component in the horny layer of the skin, that
improve reduced skin tone and cosmetic fitness. The biosurfactant
alone has ceramide-like skin roughness improvement/skin care
actions; the effects of the biosurfactant can be improved by
combining it with ceramide.
[0034] One advantage of a biosurfactant having ceramide-like
actions is that it can be easily produced on a large scale and used
as an emulsifier. Therefore, the biosurfactant of the present
invention is more versatile than existing ceramide.
[0035] As the biosurfactant, trehalose lipid, rhamnolipid,
sophorolipid, surfactin, spiculisporic acid, emulsan, MEL, MML and
the like may be used, but it is preferable to use biosurfactants
forming a lamella structure. Among these biosurfactants, MEL and
MML are preferable, MEL-A, MEL-B or MEL-C are more preferable, and
MEL-B or MEL-C are particularly preferable.
[0036] Biosurfactants forming the lamella structure include MEL and
MML, and particularly include MEL.
[0037] In one embodiment of the present invention, preferable MELs
includes three types of MEL-A represented by the formula (I), MEL-B
represented by the formula (II) and MEL-C represented by the
formula (III); these may be used alone or in combination. MEL-A
represented by the formula (I) and MEL-B represented by the formula
(II) are more preferable, and MEL-B represented by the formula (II)
is the most preferable.
[0038] MML is represented by the formula (IV) (in the formula,
either or both of the acetyl groups at positions 4 and 6 in mannose
may be substituted with hydroxyl group(s)).
[0039] R.sup.1 and R.sup.2 each have 1 to 19 carbon atoms,
preferably 1 to 17, and more preferably 7 to 15.
[0040] R.sup.1 and R.sup.2 may be the same or different, and
include C.sub.1 to C.sub.19 alkyl groups, C.sub.2 to C.sub.19
alkenyl groups, C.sub.5 to C.sub.1 alkadienyl groups and C.sub.8 to
C.sub.19 alkatrienyl groups.
[0041] Preferable groups as R.sup.1 and R.sup.2 in the formulae
(I), (II), (III) and (IV) include C.sub.1 to C.sub.19 alkyl groups
such as CH.sub.3(CH.sub.2).sub.6, CH.sub.3(CH.sub.2).sub.8,
CH.sub.3(CH.sub.2).sub.10CH.sub.3(CH.sub.2) 12,
CH.sub.3(CH.sub.2).sub.14, CH.sub.3(CH.sub.2).sub.16 and
CH.sub.3(CH.sub.2).sub.18. MEL of the formula (I), (II) or (III)
and MML of the formula (IV) in which R.sup.1 and R.sup.2 are
C.sub.1 to C.sub.19 alkyl groups can be obtained by adding the raw
material, e.g., a saturated carboxylic acid or unsaturated
monocarboxylic acid such as formic acid, acetic acid, propionic
acid, butyric acid, valeric acid, lauric acid, myristic acid,
palmitic acid, stearic acid, oleic acid and palmitoleic acid or an
ester thereof (monoalkyl esters, mono-, di-, triglyceride, or fats
and oils including these saturated fatty acids or unsaturated
monofatty acids) to the medium. When an unsaturated carboxylic acid
such as oleic acid or palmitoleic acid is used as the raw material,
the unsaturated carboxylic acid or a product during 0-oxidation
thereof is introduced, and thus the alkyl group becomes a major
component and the alkenyl group becomes a minor component as
R.sup.1 and R.sup.2. Likewise, the MEL of the formula (I), (II) or
(III) and the MML of the formula (IV) in which R.sup.1 and R.sup.2
are C.sub.2 to C.sub.19 alkenyl groups, C.sub.5 to C.sub.19
alkadienyl groups or C.sub.8 to C.sub.19 alkatrienyl groups, can be
obtained by adding the raw material, e.g., linoleic acid, linolenic
acid, arachidonic acid, EPA, DHA having two or more double bonds or
the ester thereof (monoalkyl ester, mono-, di-, triglyceride, or
fats and oils including highly unsaturated fatty acids thereof) to
the medium. For example, when linoleic acid is used as the raw
material, the alkenyl group becomes the major component and the
alkadienyl group becomes the minor component as R.sup.1 and
R.sup.2. When linolenic acid is used as the raw material, the
alkadienyl group becomes the major component and the alkenyl group
or the alkatrienyl group becomes the minor component as R.sup.1 and
R.sup.2.
[0042] These biosurfactants may be used alone, or two or more
biosurfactants may be used in combination.
##STR00001##
[0043] In the formulae (I) to (IV), R.sup.1 and R.sup.2 may be the
same or different and represent hydrogen atoms; straight or
branched C.sub.1 to C.sub.19, preferably C.sub.1 to C.sub.17, and
more preferably C.sub.7 to C.sub.15 alkyl groups; straight or
branched C.sub.2 to C.sub.19, preferably C.sub.2 to C.sub.17, and
more preferably C.sub.7 to C.sub.15 alkenyl groups; straight or
branched C.sub.5 to C.sub.19, preferably C.sub.5 to C.sub.17, and
more preferably C.sub.7 to C.sub.15 alkadienyl groups; or straight
or branched C.sub.8 to C.sub.19, preferably C.sub.8 to C.sub.17,
and more preferably C.sub.8 to C.sub.15 alkatrienyl groups.
[0044] The method for producing the biosurfactant is not
particularly limited, and a fermentation method using a
microorganism could be optionally selected. For example, MELs
(MEL-A, MEL-B and MEL-C) can be produced by culturing Pseudozyma
antarctica NBRC 10736 according to standard methods. As the
microorganism, Pseudozyma antarctica, Pseudozyma sp. and the like
can be used. It is a well-known fact that a MEL mixture can be
easily yielded from any microorganism. The MEL mixture can be
purified using silica gel chromatography to isolate MEL-A, MEL-B
and MEL-C. Pseudozyma antarctica and Pseudozyma tsukubaensis are
known as the microorganisms that produce MEL-B, and these
microorganisms may be used. Pseudozyma hubeiensis is known as the
microorganism that produces MEL-C, and this microorganism may be
used. The microorganism with an ability to produce MEL is not
particularly limited, and can be appropriately selected depending
on the purpose.
[0045] As the medium for fermentation to produce the biosurfactant,
it is possible to use a common medium composed of N sources such as
yeast extracts and peptone, C sources such as glucose and fructose,
inorganic salts such as sodium nitrate, dipotassium hydrogen
phosphate and magnesium sulfate heptahydrate, and those in which
one or two or more non-aqueous substrates, e.g., fats and oils such
as olive oil, soybean oil, sunflower oil, corn oil, canola oil and
coconut oil, and hydrocarbons such as liquid paraffin and
tetradecane, are added thereto can be used.
[0046] Fermentation conditions such as pH value, temperature and
time period can be optionally set, and a culture medium after the
fermentation can be directly used as the biosurfactant of the
present invention. Optional manipulations such as filtration,
centrifugation, extraction, purification and sterilization can be
applied to the culture medium after fermentation, if necessary, in
the range in which the essence of the present invention is not
impaired, and it is still possible to dilute, concentrate and dry
the resulting extract.
[0047] Plant fats and oils used as raw materials are not
particularly limited, and can be appropriately selected depending
on the purpose; these can include soybean oil, rapeseed oil, corn
oil, peanut oil, cotton seed oil, safflower oil, sesame oil, olive
oil and palm oil. Among them, soybean oil is particularly
preferable in terms of enhancing production efficiency (produced
amount, production speed and yield). These may be used alone or in
combinations of two or more.
[0048] The inorganic nitrogen source is not particularly limited,
and can be appropriately selected depending on the purpose;
examples include ammonium nitrate, urea, sodium nitrate, ammonium
chloride and ammonium sulfate.
[0049] The methods for collecting and purifying the biosurfactant
are not particularly limited, and can be appropriately selected
depending on the purpose; for example, the biosurfactant can be
collected by centrifuging the culture medium to collect an oil
content and extracting with an organic solvent such as ethyl
acetate to concentrate.
[0050] As the solvent for the extraction, a mixed liquid obtained
by mixing water with an organic solvent such as alcohols (e.g.,
lower alcohols such as methanol, absolute ethanol and ethanol, or
polyvalent alcohols such as propylene glycol and 1,3-butylene
glycol), ketones such as acetone, diethyl ether, dioxane,
acetonitrile, esters such as ethyl acetate, xylene, benzene, and
chloroform can be used alone or in combinations of two or more, and
those obtained by combining respective solvent extracts can also be
used.
[0051] The method for the extraction is not particularly limited.
Typically, the extraction may be performed at temperatures ranging
from ambient temperature to a boiling point of the solvent under an
atmospheric pressure. After the extraction, the extract may be
absorbed, decolorized or purified using the filtration or the ion
exchange resin to make a solution, paste, gel or powder. In many
cases, the resulting product can be directly used, but if
necessary, a purification treatment such as a deodorant treatment
or decoloration may be given thereto in the range in which the
product's effectiveness is not affected. As a deodorant procedure
and a decoloration procedure, an activated charcoal column can be
used, and an ordinary procedure generally applied to an extracted
substance can be optionally selected and performed. If necessary, a
high-purity biosurfactant can be obtained by purification using a
silica gel column.
[0052] As the biosurfactant, MEL-A, MEL-B and MEL-C are preferable,
MEL-B and MEL-C are more preferable, and MEL-B is particularly
preferable.
[0053] The biosurfactant of the present invention used as a
cosmetic for skin care/skin roughness improvement can be produced
by the fermentation of the microorganism as described above.
[0054] The amount of the biosurfactant added to the cosmetic varies
depending on the type of the cosmetic to be targeted and cannot be
defined collectively, but can be in the range in which the skin
roughness improvement/skin care actions are not impaired.
Typically, the amount is preferably 0.001 to 50% by mass, more
preferably 0.1 to 20% by mass, still more preferably 1 to 15% by
mass and particularly preferably 3 to 10% by mass relative to each
cosmetic. Here, a use form of the biosurfactant added to the
cosmetic is optional. For example, a biosurfactant extracted from a
culture medium can be directly used, or a highly purified
biosurfactant can be used, or a biosurfactant can be used after
being suspended in water or dissolved in an organic solvent such as
ethanol.
[0055] The biosurfactant may be combined in the cosmetic by
dissolution in an oil-soluble base or an oil-soluble component, and
is preferably combined in the form of a liposome in aqueous
cosmetics such as face lotions and moisturizing liquids. The
biosurfactant combined as the liposome is preferable because it is
fused to skin cells to enhance its absorbability. The method to
prepare the liposome is not particularly limited; any of the
publicly known preparation methods, such as the ethanol injection
method or the Bangham method, can be employed.
[0056] The method for producing the cosmetic of the present
invention using the biosurfactant is not particularly limited, and
the biosurfactant can be dissolved in a nonionic surfactant, lower
alcohol, polyvalent alcohol, or natural fat or oil such as olive
oil, squalane, a fatty acid or a higher alcohol.
[0057] Examples of nonionic surfactants include sorbitan fatty acid
esters (e.g., sorbitan monooleate, sorbitan monoisostearate,
sorbitan monolaurate, sorbitan monopalmitate, sorbitan
monostearate, sorbitan sesquioleate, sorbitan trioleate, diglycerol
sorbitan penta-2-ethylhexylate, diglycerol sorbitan
tetra-2-ethylhexylate); glycerine/polyglycerine fatty acids (e.g.,
mono-cottonseed oil fatty acid glycerine, glycerine monoerucate,
glycerine sesquioleate, glycerine monostearate, glycerine
.alpha.,.alpha.'-oleate pyroglutamate, glycerine monostearate malic
acid); propylene glycol fatty acid esters (e.g., monostearic acid
propylene glycol); cured castor oil derivatives; and glycerine
alkyl ether.
[0058] Examples of POE-based hydrophilic nonionic surfactants
include POE-sorbitan fatty acid esters (e.g., POE-sorbitan
monooleate, POE-sorbitan monostearate, POE-sorbitan monooleate,
POE-sorbitan tetraoleate); POE-sorbit fatty acid esters (e.g.,
POE-sorbit monolaurate, POE-sorbit monooleate, POE-sorbit
pentaoleate, POE-sorbit monostearate); POE-glycerine fatty acid
esters (e.g., POE-glycerine monostearate, POE-glycerine
monoisostearate, POE-glycerine triisostearate, POE-monooleate);
POE-fatty acid esters (e.g., POE-distearate, POE-monodioleate,
distearic acid ethylene glycol); POE-alkyl ethers (e.g., POE-lauryl
ether, POE-oleyl ether, POE-stearyl ether, POE-behenyl ether,
POE-2-octyldodecyl ether, POE-cholestanol ether); Pluronic types
(e.g., Pluronic); POE/POP-alkyl ethers (e.g., POE/POP-cetyl ether,
POE/POP-2-decyltetradecyl ether, POE/POP-monobutyl ether,
POE/POP-hydrogenated lanoline, POE/POP-glycerine ether);
tetra-POE/tetra-POP-ethylenediamine condensates (e.g., Tetronic);
POE-castor oil cured castor oil derivatives (e.g., POE-castor oil,
POE-cured castor oil, POE-cured castor oil monoisostearate,
POE-cured castor oil triisostearate, POE-cured castor oil
pyroglutamate monoisostearate diester, POE-cured castor oil
maleate); POE-beeswax lanoline derivatives (e.g., POE-sorbit
beeswax); alkanol amide (e.g., palm oil fatty acid diethanolamide,
lauric acid monoethanolamide, fatty acid isopropanolamide);
POE-propylene glycol fatty acid ester; POE-alkylamine; POE-fatty
acid amide; sucrose fatty acid ester; alkylethoxydimethylamine
oxide; and trioleyl phosphate.
[0059] Examples of lower alcohols include ethanol, propanol,
isopropanol, isobutyl alcohol and t-butyl alcohol.
[0060] Examples of polyvalent alcohols include bivalent alcohols
(e.g., ethylene glycol, propylene glycol, trimethylene glycol,
1,2-butylene glycol, 1,3-butylene glycol, tetramethylene glycol,
2,3-butylene glycol, pentamethylene glycol, 2-butene-1,4-diol,
hexylene glycol, octylene glycol); trivalent alcohols (e.g.,
glycerine, trimethylolpropane); tetravalent alcohols (e.g.,
pentaerythritols such as 1,2,6-hexanetriol); pentavalent alcohols
(e.g., xylitol); hexavalent alcohols (e.g., sorbitol, mannitol);
polyvalent alcohol polymers (e.g., diethylene glycol, dipropylene
glycol, triethylene glycol, polypropylene glycol, tetraethylene
glycol, diglycerine, polyethylene glycol, triglycerine,
tetraglycerine, polyglycerine); bivalent alcohol alkyl ethers
(e.g., ethylene glycol monomethyl ether, ethylene glycol monoethyl
ether, ethylene glycol monobutyl ether, ethylene glycol monophenyl
ether, ethylene glycol monohexyl ether, ethylene glycol
mono-2-methylhexyl ether, ethylene glycol isoamyl ether, ethylene
glycol benzyl ether, ethylene glycol isopropyl ether, ethylene
glycol dimethyl ether, ethylene glycol diethyl ether, ethylene
glycol dibutyl ether); bivalent alcohol alkyl ethers (e.g.,
diethylene glycol monomethyl ether, diethylene glycol monoethyl
ether, diethylene glycol monobutyl ether, diethylene glycol
dimethyl ether, diethylene glycol diethyl ether, diethylene glycol
butyl ether, diethylene glycol methyl ethyl ether, triethylene
glycol monomethyl ether, triethylene glycol monoethyl ether,
propylene glycol monomethyl ether, propylene glycol monoethyl
ether, propylene glycol monobutyl ether, propylene glycol isopropyl
ether, dipropylene glycol methyl ether, dipropylene glycol ethyl
ether, dipropylene glycol butyl ether); bivalent alcohol ether
esters (e.g., ethylene glycol monomethyl ether acetate, ethylene
glycol monoethyl ether acetate, ethylene glycol monobutyl ether
acetate, ethylene glycol monophenyl ether acetate, ethylene glycol
diadipate, ethylene glycol disuccinate, diethylene glycol monoethyl
ether acetate, diethylene glycol monobutyl ether acetate, propylene
glycol monomethyl ether acetate, propylene glycol monoethyl ether
acetate, propylene glycol monopropyl ether acetate, propylene
glycol monophenyl ether acetate); glycerine monoalkyl ethers (e.g.,
chimyl alcohol, selachyl alcohol, bathyl alcohol); sugars and sugar
alcohols (e.g., sorbitol, maltitol, maltotriose, mannitol, sucrose,
erythritol, glucose, fructose, starch-degraded sugar, maltose,
xylitose, starch-degraded sugar-reduced alcohol); glysolid;
tetrahydrofurfuryl alcohol; POE-tetrahydrofurfuryl alcohol;
POP-butyl ether; POP/POE-butyl ether; tripolyoxypropylene glycerine
ether; POP-glycerine ether; POP-glycerine ether phosphate;
POP/POE-pentaerythritol ether, and polyglycerine.
[0061] Examples of oils include animal and plant oils such as
avocado oil, olive oil, sesame oil, camellia oil, evening primrose
oil, turtle oil, macadamia nut oil, corn oil, mink oil, rapeseed
oil, egg yolk oil, parsic oil, wheat germ oil, sasanqua oil, castor
oil, flaxseed oil, safflower oil, cotton seed oil, perilla oil,
soybean oil, peanut oil, tea oil, kaya seed oil, rice bran oil,
China wood oil, jojoba oil, cacao butter, fractionated coconut oil,
horse oil, palm oil, palm kernel oil, beef tallow, mutton tallow,
lard, lanoline, whale wax, beeswax, carnauba wax, vegetable wax,
candelilla wax and squalane, cured oils thereof, mineral oils such
as liquid paraffin and petrolatum, and synthetic triglycerines such
as tripalmitate glycerine.
[0062] Examples of the fatty acid include lauric acid, myristic
acid, palmitic acid, oleic acid, linoleic acid, linolenic acid,
stearic acid, behenic acid, 12-hydroxystearic acid, isostearic
acid, undecynoic acid, tolic acid, eicosapentaenoic acid and
docosahexaenoic acid. Examples of the higher alcohol include lauryl
alcohol, cetyl alcohol, stearyl alcohol, behenyl alcohol, myristyl
alcohol, oleyl alcohol, cetostearyl alcohol, jojoba alcohol,
lanoline alcohol, batyl alcohol, 2-decyltetradecanol, cholesterol,
phytosterol and isostearyl alcohol. Examples of the synthetic ester
include cetyl octanoate, octyldodecyl myristate, isopropyl
myristate, myristyl myristate, isopropyl palmitate, butyl stearate,
hexyl laurate, decyl oleate, dimethyloctanoic acid, cetyl lactate
and myristyl lactate. Examples of the silicone include chain-shaped
polysiloxanes such as dimethyl polysiloxane and methylphenyl
polysiloxane, cyclic polysiloxanes such as decamethyl
cyclopolysiloxane, and three-dimensional mesh structures of
silicone resins.
[0063] The skin care cosmetics of the present invention include
milky lotions, beauty liquids, creams, lotions, skin care oils,
cleansing oils, bath oils, or facial washes, makeup removers,
shampoos and body soaps.
[0064] The MELs of the present invention, particularly MEL-A, MEL-B
and MEL-C, are more excellent than ceramide in terms of ease of use
because they are easily produced and also have an emulsification
action.
EXAMPLES
[0065] The present invention will be described in more detail in
the following Examples; however, the present invention is not
limited to these Examples.
(Method for Evaluating Action to Improve Skin Roughness)
[0066] Tissues are taken out according to the main points of the
handling manual accompanying the Test Skin LSE-002 or 003 kit
(Toyobo Co., Ltd.). A ring for assuring a drug exposure site is
adhered to an LSE tissue surface, and an aqueous solution of 0.1%
sodium dodecyl sulfate (SDS) is added into the ring, which is then
left to stand at room temperature for 5 minutes. Subsequently, SDS
is removed using an aspirator, and 3 ml of an assay medium is
sprayed using a pipette to wash the tissue. By these manipulations,
moisturizing components in the horny layer were eluted, and a dry
skin was made.
[0067] Subsequently, as test articles, each 80 .mu.L of purified
water or a cosmetic liquid (Fenatty cosmetic liquid, fatted; FANCL
Corporation) was added on the LSE (living skin equivalent) tissue
surface, which was then left to stand at room temperature for 60
minutes. Then, the test articles were aspirated and removed using
the aspirator. Subsequently, the LSE tissue was placed on an assay
tray on which an assay medium had not been placed, and incubated
for 24 hours in a CO.sub.2 incubator adjusted to a temperature of
37.degree. C. and a relative humidity of 15 to 20% RH. Then, the
LSE tissue was removed from the CO.sub.2 incubator, and 1.2 mL of a
mixed solution of the assay medium containing 0.333 g/mL of a
tetrazolium salt (MTT) reagent was added to the assay tray
according to the main points of the handling manual accompanying
the LSE-003 kit. The LSE tissue in the assay tray was incubated for
3 hours in a CO.sub.2 incubator adjusted to a temperature of
37.degree. C. and a relative humidity of 15 to 20% RH.
[0068] After the treatment with MTT, a piece of the LSE tissue was
obtained by punching out a central portion of the LSE tissue that
included a polycarbonate film using a biopsy punch of 8 mm.phi.,
then transferring it to a small test tube, and 700 .mu.L of 0.04 N
hydrochloric acid-isopropanol was added thereto to extract in a
dark place for two hours. After termination of the extraction, the
extracted solution was stirred and mixed thoroughly. Subsequently,
an absorbance at 562 nm for extracted formazan having a blue-violet
color was measured. The absorbance obtained by this method is
closely associated with the effect of improving skin roughness.
Thus, this method is effective for evaluating the skin roughness
improvement quantitatively, simply and economically.
Example 1
Production of MEL
[0069] One loop of Pseudozyma antarctica NBRC 10736 was inoculated
in a seed medium (20 mL/500 mL Sakaguchi flask) to perform an
inoculum culture. The culture was performed at 30.degree. C.
overnight. The resulting culture medium was rendered in the
inoculum. The seed medium was composed of 4% glucose, 0.3%
NaNO.sub.3, 0.02% MgSO.sub.4.H.sub.2O, 0.02% KH.sub.2PO.sub.4 and
0.1% yeast extract. The above inoculum (75 mL) was inoculated in
1.5 L (5 L-jar) of a production medium, and cultured at 30.degree.
C., 300 rpm (stirring frequency) and 0.5 L/min0 (air) using the 5
L-jar. A production medium was composed of 3% soybean oil, 0.02%
MgSO.sub.4.H.sub.2O, 0.02% KH.sub.2PO.sub.4 and 0.1% yeast extract.
The culture medium (250 mL) was centrifuged (6,500 rpm, 30 min), a
supernatant was removed, and a precipitate (microbial cells) was
collected. Ethyl acetate (50 mL) was added to the precipitate,
which was then stirred thoroughly and centrifuged (8,500 rpm, 30
min) to separate the supernatant from the precipitate. The
supernatant was concentrated using an evaporator. MEL fractions
(MEL-A, MEL-B and MEL-C) were obtained by using silica gel and
eluting with hexane:acetone=5:1 and hexane:acetone=1:2.
Example 1A
Production of MEL-B
[0070] A frozen stock (0.2 mL) of P. tsukubaensis was inoculated in
20 mL of YM seed medium in a 500 mL Sakaguchi flask, and cultured
at 26.degree. C. at 180 rpm overnight to make a seed inoculum. The
seed inoculum was inoculated again in 20 mL of YM seed medium in a
500 mL Sakaguchi flask, and cultured at 26.degree. C. at 180 rpm
overnight to make an inoculum. The inoculum (20 mL) was inoculated
in 2 L of YM medium in a 5 L jar and cultured at 26.degree. C. at
300 rpm (1/4 VVM, 0.5 L air/min) for 8 days. The culture medium was
centrifuged at 7,900 rpm at 4.degree. C. for 60 minutes to separate
the microbial cells (including MEL-B) from the supernatant. Ethyl
acetate (80 mL) was added to a microbial cell fraction, which was
then shaken to be suspended thoroughly and then centrifuged at
7,900 rpm at 4.degree. C. for 30 minutes. An equivalent amount of
brine was added to the resulting supernatant, and the mixture was
stirred to yield an ethyl acetate layer. An appropriate amount of
sodium sulfate anhydrate was added to the ethyl acetate layer,
which was then left to stand for 30 minutes and evaporated to yield
a crude product of purified MEL-B. The resulting crude product (20
g) of purified MEL-B was further purified by using a silica gel
column (200 g) and eluting with hexane/acetone to yield a purified
MEL-B product.
Example 2
[0071] Although soybean oil was used as the production material in
the production of MEL in Example 1, MEL-A, MEL-B and MEL-C are
isolated and purified using olive oil as the production material
instead, and cultured the same way as in Example 1. The MEL
fractions obtained at this time are referred to as MEL-A (OL),
MEL-B (OL) and MEL-C(OL), in order to distinguish them from the MEL
obtained in Example 1.
Example 3
[0072] Although soybean oil was used as the production material in
the production of MEL in Example 1, MEL-A, MEL-B and MEL-C are
isolated and purified using methyl myristate for the production
material instead, and cultured the same way as in Example 1. The
MEL fractions obtained at this time are referred to as MEL-A (MY),
MEL-B (MY) and MEL-C (MY), in order to distinguish them from the
MEL obtained in Example 1.
Example 4
Evaluation of MEL-A in Skin Roughness Model
[0073] A skin roughness model using a three-dimensional skin model
was made as follows. The skin roughness model was made by treating
a Test Skin (LSE-002 or 003; Toyobo Co., Ltd.) with 1% SDS to
remove the lipid components in the horny layer. The skin roughness
prevention effect was examined by adding olive oil in which MEL-A
was dissolved on the cells, leaving it to stand overnight, and
subsequently calculating cell viability using a commercially
available MTT kit. As shown in FIG. 1, the cell viability increased
in a MEL-A-concentration-dependent manner, confirming that MEL-A
acts as an alternative to ceramide. Meanwhile, the olive oil alone
exhibited no such effect.
Example 5
Evaluation of MEL-B in Skin Roughness Model
[0074] The skin roughness model using the three-dimensional skin
model was made as follows. The skin roughness model was made by
treating a Test Skin (LSE-002 or 003; Toyobo Co., Ltd.) with 1% SDS
to remove the lipid components in the horny layer. The skin
roughness prevention effect was examined by adding the olive oil in
which the MEL-B obtained in Example 1A was dissolved on the cells,
leaving it to stand overnight, and subsequently calculating the
cell viability using a commercially available MTT kit. As shown in
FIG. 4, the cell viability increased in a
MEL-B-concentration-dependent manner, confirming that MEL-B acts as
an alternative to ceramide. Meanwhile, the olive oil alone
exhibited no such effect.
Example 6
Evaluation of MEL-C in Skin Roughness Model
[0075] The skin roughness model using the three-dimensional skin
model was made as follows. The skin roughness model was made by
treating a Test Skin (LSE-002 or 003 supplied from Toyobo Co.,
Ltd.) with 1% SDS to remove the lipid components in the horny
layer. The skin roughness prevention effect was examined by adding
the olive oil in which the MEL-C obtained in Example 1 was
dissolved on the cells, leaving it to stand overnight, and
subsequently calculating the cell viability using a commercially
available MTT kit. As shown in FIG. 4, the cell viability increased
in a MEL-C-concentration-dependent manner, confirming that MEL-C
acts as an alternative to ceramide. Meanwhile, the olive oil alone
exhibited no such effect.
Example 7
Evaluation of MEL-A (OL) and MEL-A (MY) in Skin Roughness Model
[0076] Similar to Example 4, a skin roughness model using the
three-dimensional skin model was made as follows. The skin
roughness model was made by treating a Test Skin (LSE-002 or 003
supplied from Toyobo Co., Ltd.) with 1% SDS to remove the lipid
components in the horny layer. The skin roughness prevention effect
was examined by adding olive oil in which MEL-A (Example 1), MEL-A
(OL) (Example 2) or MEL-A (MY) (Example 3) was dissolved on the
cells, leaving it to stand overnight, and subsequently calculating
the cell viability using a commercially available MTT kit. As shown
in FIG. 3, the cell viability increased in a
MEL-A-concentration-dependent manner, confirming that MEL-A acts as
an alternative to ceramide. Meanwhile, the olive oil alone
exhibited no such effect.
Example 8
Effect of MEL-A-Containing Cream in Human Skin Roughness Test
[0077] The skin was roughened by placing an inner side of a human's
upper arm in contact with a solution of 1% SDS for 10 minutes.
Immediately following, the above cream containing 5% MEL-A was
applied, and after 3 hours, the skin was washed with warm water.
The oil content was wiped away with a Kim towel, and the moisture
content of the skin's horny layer was measured using Skicon. As
shown in FIG. 2, the recovery of the moisture content was observed
when the MEL-A-containing cream was applied.
Example 9
Effect of MEL-B-Containing Cream in Human Skin Roughness Test
[0078] The skin was roughened by placing an inner side of a human's
upper arm in contact with a solution of 1% SDS for 10 minutes.
Immediately following, the above cream containing 5% MEL was
applied, and after 3 hours, the skin was washed with warm water.
The oil content was wiped away with Kim towel, and the moisture
content of the skin's horny layer was measured using Skicon. As
shown in FIG. 5, the recovery of the moisture content was observed
when the MEL-B- or C-containing cream was applied.
Example 10
Dispersion Stability Test of MEL-B and MEL-C
[0079] MEL-B or MEL-C at a concentration of 10 mg/mL was added to
water, stirred and suspended. Its absorbance at 650 nm was
measured. The results are shown in FIG. 6. From the results in FIG.
6, it is clear that MEL-B and MEL-C are particularly excellent in
dispersion stability.
Example 11
Preparation of a Mel-B Liposome Solution
[0080] A MEL-B liposome solution was prepared as follows using an
ethanol injection method. 10 mg of MEL-B was dissolved in 0.5 mL of
ethanol, and 1 mL of distilled water previously warmed to about
70.degree. C. was added. The mixture was slightly shaken and mixed,
and residual ethanol was distilled off using a rotary evaporator.
Sonication for about 5 minutes was applied thereto using a water
bath-type sonicator (W-220R; Honda Electronics Co., Ltd.), and then
the distilled water was added to make a total volume 1 mL.
[0081] A MEL-B liposome solution was prepared as follows using the
Bangham method. 10 mg of MEL-B was dissolved in 1 mL of chloroform,
and the solvent was distilled off using a rotary evaporator to make
a thin film. Thereto, 1 mL of distilled water was added, and
sonication for about 5 minutes was applied thereto using a water
bath-type sonicator (W-220R; Honda Electronics Co., Ltd.).
[0082] A MEL-B suspension was prepared as follows. 1 mL of
distilled water was added to 10 mg of MEL-B, and stirred using a
vortex mixer to prepare the suspension.
[0083] The MEL-B liposome aqueous solutions prepared by the ethanol
injection method, Bangham method and MEL-B suspension were
subjected to a skin roughness test using the three-dimensional skin
model shown in Example 4, and their effects were examined. The
results are shown in FIG. 7. As a result, the MEL-B liposome
aqueous solution prepared by any of the ethanol injection method,
Bangham method and the suspension exhibited the same skin roughness
improvement effect as that of the MEL-B olive oil solution (MEL-B
concentration: 1%).
Example 12
Production of Beauty Liquid
[0084] A beauty liquid having the composition shown below was
produced using standard methods. As a control, a MEL-free beauty
liquid was also produced using standard methods.
TABLE-US-00001 (Composition) (% by weight) Sorbit 4.0 Dipropylene
glycol 6.0 Polyethylene glycol 1500 5.0 POE (20) Oleyl alcohol
ether 0.5 Sucrose fatty acid ester 0.2 methyl cellulose 0.2 MEL-B
5.0 Purified water Amount to make the total 100%
Example 13
Production of Milky Lotion
[0085] A milky lotion having the composition shown below was
produced using standard methods. As the control, a MEL-free milky
lotion was also produced using standard methods.
TABLE-US-00002 (Composition) (% by weight) Glyceryl ether 1.5
Sucrose fatty acid ester 1.5 Sorbitan monostearate 1.0 Squalane 7.5
Dipropylene glycol 5.0 MEL-B 5.0 Purified water Amount to make the
total 100%
Example 14
Production of Cream
[0086] A cream having the composition shown below was produced
using standard methods. As the control, a MEL-free cream was also
produced using standard methods.
TABLE-US-00003 (Composition) (% by weight) Propylene glycol 6.0
Dibutyl phthalate 19.0 Stearic acid 5.0 Glycerine monostearate 5.0
Sorbitan monostearate 12.0 Polyethylene sorbitan monostearate 38.0
Sodium edetate 0.03 MEL-B 5.0 Purified water Amount to make the
total 100%
Example 15
Production of Oil for Makeup
TABLE-US-00004 [0087] (Composition) (% by weight) Olive oil 90
MEL-A 10
Example 16
Skin Care Oil
TABLE-US-00005 [0088] (Composition) (% by weight) Olive oil 50
MEL-C 30 Squalane 10 Sesame oil 10
Example 17
Skin Care Oil
TABLE-US-00006 [0089] (Composition) (% by weight) Olive oil 39
MEL-C 59 Sesame oil 1 Lavender oil 0.4 Rosemary oil 0.4 Sage oil
0.1 .delta.-Tocopherol 0.1
Example 18
Cleansing Oil
TABLE-US-00007 [0090] (Composition) (% by weight) Olive oil 40
MEL-B 28 Methylphenyl polysiloxane 2 Ethanol 0.3 Isostearic acid
0.1 Cetyl 2-ethylhexanoate 20 Polyethylene glycol diisostearate 2
Palm oil fatty acid diethanolamide 0.1 Polyethylene glycol
monoisostearate 2 .delta.-Tocopherol 0.1 Purified water 1 Perfume
Appropriate quantity
Example 19
Bath Oil
TABLE-US-00008 [0091] (Composition) (% by weight) Olive oil 25
MEL-A 25 Liquid paraffin 25 Neopentyl glycol dicaprylate 10
Polyoxyethylene oleyl ether 10 Purified water 0.5
.delta.-Tocopherol 0.1 Perfume Appropriate quantity
INDUSTRIAL APPLICABILITY
[0092] According to the present invention, it is possible to
prevent skin roughness by supplementing removed ceramide with the
MEL-A, MEL-B or MEL-C of the present invention, which can not only
be used as emulsifiers, but can also be used as alternatives
ceramide, as they are much more easily produced than ceramide and
are thus expected to contribute greatly to the industry.
* * * * *